The detection of androstenedione abuse in sport: a mass spectrometry strategy to identify the 4-hydroxyandrostenedione metabolite

Rapid Commun Mass Spectrom. 2008 Dec;22(24):4147-57. doi: 10.1002/rcm.3826.


Studies have shown that the administration of androstenedione (ADIONE) significantly increases the urinary ratio of testosterone glucuronide to epitestosterone glucuronide (T/E) - measured by gas chromatography/mass spectrometry (GC/MS) - in subjects with a normal ( approximately 1) or naturally high (>1) initial values. However, the urinary T/E ratio has been shown not to increase in subjects with naturally low (<1) initial values. Such cases then rely on the detection of C(6)-hydroxylated metabolites shown to be indicative of ADIONE administration. While these markers may be measured in the routine GC/MS steroid profile, their relatively low urinary excretion limits the use of gas chromatography/combustion/isotope ratio mass spectrometry (GC/C/IRMS) to specifically confirm ADIONE administration based on depleted (13)C content. A mass spectrometry strategy was used in this study to identify metabolites of ADIONE with the potential to provide compound-specific detection. C(4)-hydroxylation was subsequently shown to be a major metabolic pathway following ADIONE administration, thereby resulting in urinary excretion of 4-hydroxyandrostenedione (4OH-ADIONE). Complementary analysis of 4OH-ADIONE by GC/MS and GC/C/IRMS was used to confirm ADIONE administration.

MeSH terms

  • Adult
  • Androstenedione / analogs & derivatives*
  • Androstenedione / urine*
  • Chromatography, Gas / methods
  • Chromatography, High Pressure Liquid
  • Doping in Sports*
  • Humans
  • Male
  • Mass Screening / methods
  • Reproducibility of Results
  • Spectrometry, Mass, Electrospray Ionization / methods*
  • Spectrophotometry, Infrared
  • Substance Abuse Detection / methods*
  • Young Adult


  • Androstenedione
  • formestane